Proliferation and differentiation of pancreatic ?-cells: ultrastructural analysis of the pancreas in diabetic mice induced by selective alloxan perfusion

Abstract

To clarify the mechanism of regenerative processes of pancreatic β-cells, we constructed a new diabetic model of mice and investigated their pancreatic endocrine cells by electron microscopy. Male ICR mice (8 weeks old) were partially and chemically depancreatized by perfusing alloxan (100 mg/kg body weight) via the caudal vein after clamping the cranial mesenteric artery. By this method, we could render the mice diabetic by partial reduction of β-cells localized in the splenic, gastric, and parabiliary segment. Glucose intolerance gradually ameliorated without any treatment. In the perfused segments, pancreatic β-cells showed pyknosis and the mitochondria were swollen 6h after the treatment, while non-β-cells including α-cells remained intact. At 5 days, β-cells were few and the islets became smaller in size. At 20 weeks, small islet cell clusters (ICCs) were observed budding from interlobular and intralobular ductal epithelial cells. β-cells scattering in the exocrine pancreas were also frequently observed. In the alloxan-nonperfused segment, β-cells with thin rough endoplasmic reticulum and immature secretory granules without an electron-opaque halo were observed, and the number of mitochondria increased in some β-cells at 1 day and 5 days after the treatment. At 20 weeks, β-cells that contained only mature granules were observed in hypertrophic islets. In this model, both proliferation of residual β-cells and differentiation of pancreatic endocrine cells from the ductal epithelial cells were recognized.